Genetic Analysis of the Cronobacter sakazakii O4 to O7 O-Antigen Gene Clusters and Development of a PCR Assay for Identification of All C. sakazakii O Serotypes
Genetic Analysis of the Cronobacter sakazakii O4 to O7 O-Antigen Gene Clusters and Development of a PCR Assay for Identification of All C. sakazakii O Serotypes
复制标题
坂崎克罗诺杆菌 O4 至 O7 O 抗原基因簇的遗传分析以及用于鉴定所有坂崎克罗诺杆菌 O 血清型的 PCR 检测方法的开发
DOI：
10.1128/aem.07825-11
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发表时间：
2012-06-01
影响因子：
4.4
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Sun, Yamin;Wang, Min;Wang, Lei
革兰氏阴性菌坂崎克罗诺杆菌是一种新兴的食源性病原体，可引起新生儿严重的侵袭性感染。外膜o抗原脂多糖的变化为革兰氏阴性菌的血清分型提供了依据。最近建立了7种血清型（O1 ~ O7）阪崎弧菌o抗原分型方案，并对3种血清型（O1、O2和O3）的o抗原基因簇和特异性引物进行了鉴定。本研究对阪崎c O4、O5、O6和O7 o抗原基因簇进行了测序，并在同源性的基础上对基因功能进行了预测。阪崎C. O4与大肠杆菌O103具有相似的o抗原基因簇。分析了7个坂崎梭菌o -抗原基因簇的一般特征和异常，并探讨了o -抗原结构与其基因簇之间的关系。通过对136株阪崎曲菌及其近缘种的筛选，确定了O4 ~ O7的血清型特异性基因，并建立了所有阪崎曲菌O血清型的多重PCR分型方法。pcr血清分型方法的灵敏度为0.01 ng基因组DNA和103 CFU /ml。本研究完成了阪崎弧菌o抗原遗传学的阐明，为阪崎弧菌O1 ~ O7菌株的鉴定提供了一种适合的分子方法。
ABSTRACT The Gram-negative bacterium Cronobacter sakazakii is an emerging food-borne pathogen that causes severe invasive infections in neonates. Variation in the O-antigen lipopolysaccharide in the outer membrane provides the basis for Gram-negative bacteria serotyping. The O-antigen serotyping scheme for C. sakazakii, which includes seven serotypes (O1 to O7), has been recently established, and the O-antigen gene clusters and specific primers for three C. sakazakii serotypes (O1, O2, and O3) have been characterized. In this study, the C. sakazakii O4, O5, O6, and O7 O-antigen gene clusters were sequenced, and gene functions were predicted on the basis of homology. C. sakazakii O4 shared a similar O-antigen gene cluster with Escherichia coli O103. The general features and anomalies of all seven C. sakazakii O-antigen gene clusters were evaluated and the relationship between O-antigen structures and their gene clusters were investigated. Serotype-specific genes for O4 to O7 were identified, and a molecular serotyping method for all C. sakazakii O serotypes, a multiplex PCR assay, was developed by screening against 136 strains of C. sakazakii and closely related species. The sensitivity of PCR-based serotyping method was determined to be 0.01 ng of genomic DNA and 103 CFU of each strain/ml. This study completes the elucidation of C. sakazakii O-antigen genetics and provides a molecular method suitable for the identification of C. sakazakii O1 to O7 strains.